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Ray
On Thu, 1 Feb 1996, Weaverli wrote:
> Help, anyone!
>
> One of my flow "customers" is growing a rat mammary adenocarcinoma
> cell line, MAT B III (which appears in ATCC's repository as CRL 1666)
> for in vitro studies. The cells grow as a static suspension, but in
> aggregates of about 5-10 cells, sometimes more. The problem with
> these cells is not initially dispersing the aggregates, but having
> them remain dispersed. The group has tried an assortment of agents or
> combinations thereof, including trypsin, trypsin-EDTA, EDTA,
> collagenase, DNAse, and hyaluronidase (followed by appropriate
> inhibitors or washing steps to remove enzyme activity.) The
> aggregates disperse, but then re-aggregate, even when placed on ice.
> This results in my turning away whole experiments consisting of clumps
> of very healthy, intact cell aggregates (100% of the cells
> re-aggregate...not a single cell can be found). Mechanical disruption
> produces no single cells.
>
> Is or has ANYONE worked with MAT B III? If so, have you had any
> success with keeping them dispersed for flow analysis? Is it time to
> turn to microscopy/image analysis, or throw in the towel and find
> another cell line? Your comments are appreciated!
>
> Linda Weaver
>
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